The first lane shows the DNA size standard (DNA-Ladder). Sample 1 is positive for GA733-2, Muc-1 and CA125, samples 3, 4 and 5 are positive for GA733-2 and Muc-1, and sample 6 is positive for GA733-2. Sample 2 is negative. Actin is detected in samples 1 to 6. The PCR negative (C-) and positive control are shown in the last two lanes.

AdnaTest OvarianCancerDetect results of duplex-PCR samples

The first lane shows the DNA size standard (DNA-Ladder). Samples 1 and 3 are positive for ERCC1. Sample 2 is negative. Actin is detected in samples 1 to 3. The PCR negative (C-) and positive control (ERCC1) are shown in the last two lanes.

AdnaTest workflow

AdnaTest OvarianCancerSelect enables the immunomagnetic
enrichment of tumor cells via epithelial and tumor-associated antigens. Antibodies against
epithelial and tumor-associated antigens are conjugated to magnetic beads (Dynabeads) for
labeling of tumor cells in peripheral blood. Labeled cells are extracted by a magnetic particle
concentrator (AdnaMag-L and AdnaMag-S) and are subsequently lysed. The results of the
AdnaTest are generated within 8 hours.

In the first step, the CTCs in the blood are enriched (AdnaTest Select).
This is achieved using antibody-coated magnetic particles (beads). Several antibodies are used,
which bind with high specificity and affinity to the corresponding cancer cells. The enriched cells
are lysed and subsequently purified several times to extract mRNA.

In a second step the enriched cells are examined by RT-PCR for tumor-associated expression
patterns. The mRNA strands are reverse transcribed into cDNA. Subsequently, several tumor-associated markers can be amplified using multiplex PCR and visualized.

CE-IVD marked.

In vitro diagnostic medical device.

Performance

AdnaTest OvarianCancer is a highly specific immunomagnetic cell selection system for enriching circulating tumor cells from peripheral blood and includes a highly sensitive RT-PCR for detecting ovarian cancer-specific mRNA markers.

Principle

AdnaTest technology uses an optimized combination of antibodies for cell selection in 5 ml whole blood and a combination of tumor-associated markers for the detection of tumor cells: EpCAM, Muc1, CA125 and ERCC1.

Procedure

The AdnaTest uses a two-step process (Select and Detect) to generate results within 5 hours (see figures “AdnaTest workflow”). AdnaTest OvarianCancerSelect enables the immunomagnetic enrichment of tumor cells from whole blood via epithelial- and tumor-associated antigens. Antibodies against epithelial- and tumor-associated antigens are conjugated to magnetic beads for labeling of tumor cells. Labeled cells are extracted by a magnetic particle concentrator (AdnaMag-L and AdnaMag-S), lysed and mRNA is then purified from these lysates (see figures “AdnaTest Select – Immunomagnetic cell selection with multiple tumor-associated antibodies labeled to magnetic beads”).
In the second step, the isolated mRNA is transcribed into cDNA and can be amplified using multiplex PCR (see figures “AdnaTest Detect – Multiplex PCR of various cancer-associated tumor markers”). AdnaTest OvarianCancerDetect contains Oligo (dT) 25-coated beads for the isolation of mRNA from the lysate of pre-enriched tumor cells. Reverse transcription results in cDNA that can subsequently be used as template for tumor cell detection and characterization by multiplex PCR. Multiplex PCR provides tumor-associated gene expression profiles for a variety of relevant tumor markers to ensure that the selected cells are cancer cells. The PrimerMix OvarianDetect allows the amplification of three tumor associated antigens and one control gene. The PrimerMix ERCC1-Detect amplifies the excision repair cross-complementing 1 gene (ERCC1) and one control gene. The two primer-mixes generate the following fragments: